Salt of pyrrolo[2,3-D]pyrimidine compound and novel polymorph of salt

ABSTRACT

The present invention relates to novel crystalline forms of 7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-car boxylic acid dimethylamide(compound I), its salts, and process for preparation thereof. Crystalline forms in the present invention have good stability, low hygroscopicity, good processability, easy treatability and other favorable properties. In addition, the process is simple, low cost, and has an important value for future optimization and development of the drug.

TECHNICAL FIELD

The present invention relates to the field of pharmaceutical, especiallysalts of pyrrolo[2,3-d]pyrimidine and crystalline forms thereof.

BACKGROUND OF THE INVENTION

Cyclin-dependent kinase 4 and 6 (CDK4/6) are a group of serine/threoninekinases that drive cells through G1 into S phase by associating withcyclin D. The “cyclin D-CDK4/6-INK4-Rb pathway” is universally disruptedin human cancer and the alterations accelerating of G1 progressionprovides a survival advantage to cancer cells. Then, inhibition of CDK4and CDK6 kinase activity may be a useful anticancer treatment.

LEE011 is a small molecule CDK4/6 inhibitor. It is developed by Novartisfor the treatment of breast cancer and melanoma. The clinical form ofLEE011 is succinate. LEE011 is currently in Phase III studies, and thePhase III shows positive data. The chemical name of LEE011 is7-cyclopentyl-2-(5-piperazin-1-yl-pyridin-2-ylamino)-7H-pyrrolo[2,3-d]pyrimidine-6-carboxylic acid dimethyl amide, and it has the structural of formula I:

At present, CN103201275A reports a hydrate form and a non-hydrate formof compound I mono-succinate. The hydrate form has poor solubility,lower than 0.5 mg/mL in water. The non-hydrate form has bettersolubility. The inventor of the present invention discovered that themono-succinate of the prior art is not stable and will have crystaltransformation in high humidity, and it is not good enough fordevelopment and storage. At present, there is no other crystalline formof compound I and its salts.

Based on the situation, it is necessary to develop new crystalline formswhich has good stability, low hygroscopicity, and is suitable forstorage and industrial process. And the new forms should meet therequirements of further drug development.

SUMMARY OF THE INVENTION

The present invention provides salts of compound I, its crystallineforms and preparation process, which are suitable for pharmaceuticaldevelopment and industrial process.

An objective of the present invention is to provide a hemi-succinate ofcompound I.

Further, the hemi-succinate provided by the present invention is incrystalline form, abbreviated as Form A.

Specifically, the X-ray powder diffraction pattern of the crystallineForm A shows characteristic peaks at 2theta values of 23.9°±0.2°,20.0°±0.2°, 22.1°±0.2°.

Further, the X-ray powder diffraction pattern of the crystalline Form Ashows one or two or three of the characteristic peaks at 2theta valuesof 22.0°±0.2°, 21.3°±0.2°, 13.0°±0.2°. Preferably, Form A showscharacteristic peaks at 22.0°±0.2°, 21.3°±0.2°, 13.0°±0.2°.

Further, the X-ray powder diffraction pattern of the crystalline Form Ashows one or two or three of the characteristic peaks at 2theta valuesof 4.7°±0.2°, 14.2°±0.2°, 10.6°±0.2°. Preferably, Form A showscharacteristic peaks at 4.7°±0.2°, 14.2°±0.2°, 10.6°±0.2°.

Further, the X-ray powder diffraction pattern of the crystalline Form Aof the present invention is substantially as shown in FIG. 1.

Another objective of the present invention is to provide a process forpreparing crystalline Form A of hemi-succinate, which comprises: addingcompound I and succinic acid or compound I mono-succinate in alcohols,ketones, ethers, esters or a mixture of alcohols or ketones or ethers oresters with water, stirring until solids precipitate out.

Further, said alcohol is ethanol, said ketone is acetone, said ester isethyl acetate, said ether is tetrahydrofuran.

Compared with the prior art, firstly, hemi-succinate Form A is morephysically stable, particularly it is stable at various humidity.Secondly, the mono-succinate prepared by prior art will easily havecrystal transformation in various solvents, and has poor reproducibilityin process development.

Another objective of present invention is to provide a pharmaceuticalcomposition comprising a therapeutically effective amount ofhemi-succinate Form A and pharmaceutical acceptable carrier. Generallythe therapeutically effective amount of hemi-succinate Form A is mixedor contacted with one or more pharmaceutical excipients to preparepharmaceutical composition or formulation. The pharmaceuticalcomposition or formulation is prepared by well-known methods inpharmaceutical field.

The pharmaceutical composition of hemi-succinate Form A can be used forpreparing drugs in the treatment of cancer, especially for preparingdrugs in the treatment of breast cancer and melanoma.

Another objective of the present invention is to provide a newcrystalline form of compound I mono-succinate, abbreviated as Form I,which has good stability, low hygroscopicity, and is suitable forstorage and industrial process.

Particularly, the X-ray powder diffraction pattern of the crystallineForm I of the present invention shows characteristic peaks at 2thetavalues of 11.9°±0.2°, 19.4°±0.2°, 20.6°±0.2°.

Further, the X-ray powder diffraction pattern of the crystalline Form Ishows one or two or three of the characteristic peaks at 2theta valuesof 22.7°±0.2°, 24.4°±0.2°, 26.3°±0.2°. Preferably, Form I of the presentinvention shows characteristic peaks at 22.7°±0.2°, 24.4°±0.2°,26.3°±0.2°.

Further, the X-ray powder diffraction pattern of the crystalline Form Ishows one or two or three of the characteristic peaks at 2theta valuesof 7.8°±0.2°, 15.7°±0.2°, 16.7°±0.2°. Preferably, Form I showscharacteristic peaks at 7.8°±0.2°, 15.7°±0.2°, 16.7°±0.2°.

Further, the X-ray powder diffraction pattern of the crystalline Form Iof the present invention is substantially as shown in FIG. 5.

Form I of mono-succinate of the present invention is an anhydrate. Thedifferential scanning calorimetry analysis curve (DSC) of crystallineForm I shows an endothermic peak around 197° C. (onset temperature),which is substantially as shown in FIG. 7.

The thermal gravimetric analysis (TGA) thermogram of crystalline Form Iof the present invention shows about 2.0% weight loss up to 178° C.,which is substantially as shown in FIG. 8.

Another objective of present invention is to provide a process forpreparing crystalline Form I of compound I mono-succinate, whichcomprises: dissolving compound I mono-succinate in a mixture of alcoholswith one or more solvent selected from alkyl nitriles, alkanes, ordissolving compound I mono-succinate in a mixture of alkyl nitriles withwater, stirring until solids precipitate out.

Preferably, said alcohols are methanol and ethanol, or combinationthereof, said alkyl nitrile is acetonitrile, said alkane is n-heptane.Specifically, said mixture are acetonitrile and methanol, or ethanol andn-heptane, or acetonitrile and water.

Another objective of present invention is to provide a pharmaceuticalcomposition comprising a therapeutically effective amount ofmono-succinate Form I and pharmaceutical acceptable carrier. Generallythe therapeutically effective amount of mono-succinate Form I is mixedor contacted with one or more pharmaceutical excipients to preparepharmaceutical composition or formulation. The pharmaceuticalcomposition or formulation is prepared by well-known methods inpharmaceutical field.

Further, the pharmaceutical composition of mono-succinate Form I can beused for preparing drugs in the treatment of cancer, especially forpreparing drugs in the treatment of breast cancer and melanoma.

Compared with prior art, mono-succinate Form I of the present inventionis more physically stable. Specifically, firstly, the mono-succinateprepared by prior art will convert to Form I of the present invention atvarious temperature in specific solvent system. Secondly, Form I of thepresent invention is stable at high humidity, while the mono-succinateprepared by prior art will have crystal transformation at high humidity.

The mono-succinate Form I of the present invention has lowhygroscopicity and no particular drying condition is required inpreparation. Thus it simplifies preparation and post treatment process,and suitable for industrial process. The mono-succinate Form I of thepresent invention does not need a special storage condition, thus itlowers the cost of the storage and quality control.

Another objective of present invention is to provide a pharmaceuticalcomposition comprising a therapeutically effective amount ofhemi-succinate Form A or mono-succinate Form I or combination thereofand pharmaceutical acceptable carrier. Generally the therapeuticallyeffective amount of hemi-succinate Form A or mono-succinate Form I orcombination thereof is mixed or contacted with one or morepharmaceutical excipients to prepare pharmaceutical composition orformulation. The pharmaceutical composition or formulation is preparedby well-known methods in pharmaceutical field.

Further, the pharmaceutical composition of hemi-succinate Form A ormono-succinate Form I or combination thereof can be used for preparingdrugs in the treatment of cancer, especially for preparing drugs in thetreatment of breast cancer and melanoma.

Another objective of the present invention is to provide salts ofcompound I, comprising adipate, maleate and glycollate.

Further, the adipate of compound I of the present invention is incrystalline form, abbreviated as adipate Form A.

Specifically, the X-ray powder diffraction pattern of the adipate Form Aof the present invention shows characteristic peaks at 2theta values of22.2°±0.2°, 19.2°±0.2°.

Further, the X-ray powder diffraction pattern of the adipate Form Ashows characteristic peaks at 2theta values of 24.9°±0.2°, 14.0°±0.2°,16.1°±0.2°.

Further, the X-ray powder diffraction pattern of the adipate Form Ashows one or two or three of the characteristic peaks at 2theta valuesof 18.0°±0.2°, 19.8°±0.2°, 4.8°±0.2°. Preferably, adipate Form A showscharacteristic peaks at 18.0°±0.2°, 19.8°±0.2°, 4.8°±0.2°.

Further, the X-ray powder diffraction pattern of the adipate Form A issubstantially as shown in FIG. 13.

The differential scanning calorimetry analysis curve (DSC) of adipateForm A of the present invention shows an endothermic peak around 177° C.(onset temperature), which is substantially as shown in FIG. 14.

The thermal gravimetric analysis (TGA) thermogram of adipate Form A ofthe present invention shows about 2.1% weight loss up to 159° C., whichis substantially as shown in FIG. 15.

Further, the maleate of compound I of the present invention is incrystalline form, abbreviated as maleate Form A.

Specifically, the X-ray powder diffraction pattern of the maleate Form Aof the present invention shows characteristic peaks at 2theta values of18.6°±0.2°, 19.9°±0.2°, 14.9°±0.2°.

Further, the X-ray powder diffraction pattern of the maleate Form Ashows one or two or three of the characteristic peaks at 2theta valuesof 24.5°±0.2°, 17.1°±0.2°, 16.5°±0.2°. Preferably, maleate Form A ofthis invention shows characteristic peaks at 24.5°±0.2°, 17.1°±0.2°,16.5°±0.2°.

Further, the X-ray powder diffraction pattern of the maleate Form Ashows one or two or three of the characteristic peaks at 2theta valuesof 21.9°±0.2°, 29.3°±0.2°, 8.5°±0.2°. Preferably, maleate Form A of thisinvention shows characteristic peaks at 21.9°±0.2°, 29.3°±0.2°,8.5°±0.2°.

Further, the X-ray powder diffraction pattern of the maleate Form A ofthe present invention is substantially as shown in FIG. 16.

The differential scanning calorimetry analysis curve (DSC) of maleateForm A of the present invention shows an endothermic peak around 207° C.(onset temperature), which is substantially as shown in FIG. 17.

The thermal gravimetric analysis (TGA) thermogram of maleate Form A ofthe present invention shows about 3.1% weight loss up to 138° C., whichis substantially as shown in FIG. 18.

Further, the glycollate of compound I of the present invention is incrystalline form, abbreviated as glycollate Form A.

Specifically, the X-ray powder diffraction pattern of the glycollateForm A of the present invention shows characteristic peaks at 2thetavalues of 21.3°±0.2°, 19.5°±0.2°, 23.3°±0.2°.

Further, the X-ray powder diffraction pattern of the glycollate Form Ashows one or two or three of the characteristic peaks at 2theta valuesof 21.8°±0.2°, 12.4°±0.2°, 10.1°±0.2°. Preferably, glycollate Form A ofthis invention shows characteristic peaks at 21.8°±0.2°, 12.4°±0.2°,10.1°±0.2°.

Further, the X-ray powder diffraction pattern of the glycollate Form Ashows one or two or three of the characteristic peaks at 2theta valuesof 13.3°±0.2°, 16.8°±0.2°, 23.9°±0.2°. Preferably, glycollate Form A ofthis invention shows characteristic peaks at 13.3°±0.2°, 16.8°±0.2°,23.9°±0.2°.

Further, the X-ray powder diffraction pattern of the glycollate Form Aof the present invention is substantially as shown in FIG. 19.

The differential scanning calorimetry analysis curve (DSC) of glycollateForm A of the present invention shows an endothermic peak at 253° C.(onset temperature), which is substantially as shown in FIG. 20.

The thermal gravimetric analysis (TGA) of glycollate Form A of thepresent invention shows about 3.5% weight loss up to 176° C., which issubstantially as shown in FIG. 21.

Another objective of the present invention is to provide a process forpreparing adipate Form A, comprising: adding compound I and adipic acidin alcohols, ketones, or a mixture of alcohols or ketones with water,stirring until solids precipitate out.

Further, said alcohol is ethanol, said ketone is acetone.

Further, the volume ratio of said mixture of alcohols or ketones withwater is 10:1 to 20:1.

Further, the mole ratio of compound I and adipic acid is 1:1˜1:1.2.

Another objective of the present invention is to provide a process forpreparing maleate Form A, which comprises: adding compound I and maleicacid in a mixture of ketones with water, stirring until solidsprecipitate.

Further, said ketone is acetone.

Further, the volume ratio of said ketone and water is 10:1 to 20:1.

Further, the volume ratio of said ketone and water is 19:1.

Further, the mole ratio of compound I and maleic acid is 1:0.9˜1:1.2.

Another objective of the present invention is to provide a process forpreparing glycollate Form A, comprising: adding compound I and glycolicacid in ketones, ethers or a mixture of ketones or ethers with water,stirring until solids precipitate out.

Further, said ketone is acetone, said ether is tetrahydrofuran.

Further, the volume ratio of said mixture of ketones or esters withwater is 10:1 to 20:1.

Further, the mole ratio of compound I and glycolic acid is 1:1˜1:4.5.

The crystalline adipate, maleate, glycollate of the present inventionhas low hygroscopicity and no particular drying condition is required inpreparation, thus it simplifies preparation and post treatment process,and is suitable for industrial process. The water content of these threecrystalline salts remain constant and does not need a special storagecondition, thus it lowers the cost of storage and quality control.Compared with the mono-succinate non-hydrate form in CN103201275A, thesethree crystalline salts of the present invention are more stable. Theywill not easily have crystal transformation during storage, thus havegreat economic value.

Another objective of present invention is to provide a pharmaceuticalcomposition comprising a therapeutically effective amount of crystallineadipate, maleate, glycollate of the present invention and pharmaceuticalacceptable carrier. Generally the therapeutically effective amount ofcrystalline adipate, maleate, glycollate is mixed or contacted with oneor more pharmaceutical excipients to prepare pharmaceutical compositionor formulation. The pharmaceutical composition or formulation isprepared by well-known methods in pharmaceutical field.

Further, the pharmaceutical composition of crystalline adipate, maleate,glycollate of the present invention can be used for preparing drugs inthe treatment of cancer, especially for preparing drugs in the treatmentof breast cancer and melanoma.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 XRPD pattern of hemi-succinate Form A;

FIG. 2 ¹H NMR spectrum of hemi-succinate Form A;

FIG. 3 DVS curve of hemi-succinate Form A;

FIG. 4 XRPD overlay of hemi-succinate Form A before and after DVS (thepattern above is XRPD pattern of Form A before test, the pattern belowis XRPD pattern of Form A after test);

FIG. 5 XRPD pattern of mono-succinate Form I;

FIG. 6 ¹H NMR spectrum of mono-succinate Form I;

FIG. 7 DSC curve of mono-succinate Form I;

FIG. 8 TGA curve of mono-succinate Form I;

FIG. 9 DVS curve of mono-succinate Form I (0-95% RH cycle);

FIG. 10 XRPD overlay of mono-succinate Form I before and afterhygroscopicity test (the pattern above is XRPD pattern of Form I beforetest, the pattern below is XRPD pattern of Form I after test);

FIG. 11 DVS curve of mono-succinate non-hydrate form in CN103201275A(0-95% RH cycle);

FIG. 12 XRPD overlay of mono-succinate non-hydrate form in patentCN103201275A before and after hygroscopicity test (the pattern above isXRPD pattern of mono-succinate in CN103201275A before test, the patternbelow is XRPD pattern of mono-succinate in CN103201275A after test);

FIG. 13 XRPD pattern of adipate Form A;

FIG. 14 DSC curve of adipate Form A;

FIG. 15 TGA curve of adipate Form A;

FIG. 16 XRPD pattern of maleate Form A;

FIG. 17 DSC curve of maleate Form A;

FIG. 18 TGA curve of maleate Form A;

FIG. 19 XRPD pattern of glycollate Form A;

FIG. 20 DSC curve of glycollate Form A;

FIG. 21 TGA curve of glycollate Form A;

FIG. 22 ¹H NMR spectrum of adipate Form A;

FIG. 23 ¹H NMR spectrum of maleate Form A;

FIG. 24 ¹H NMR spectrum of glycollate Form A;

FIG. 25 XRPD overlay of the stability of adipate Form A (a is the XRPDpattern of initial sample, b is XRPD pattern of adipate Form A placingat 25° C./60% RH for 30 days, c is XRPD pattern of adipate Form Aplacing at 40° C./75% RH for 30 days);

FIG. 26 XRPD overlay of the stability of maleate Form A (a is the XRPDpattern of initial sample, b is XRPD pattern of maleate Form A placingat 25° C./60% RH for 30 days, c is XRPD pattern of maleate Form Aplacing at 40° C./75% RH for 30 days);

FIG. 27 XRPD overlay of the stability of glycollate Form A (a is theXRPD pattern of initial sample, b is XRPD pattern of glycollate Form Aplacing at 25° C./60% RH for 30 days, c is XRPD pattern of glycollateForm A placing at 40° C./75% RH for 30 days);

FIG. 28 DVS curve of adipate Form A (0-95% RH cycle);

FIG. 29 XRPD overlay of adipate Form A before and after hygroscopicitytest (a is XRPD pattern of adipate Form A before test, b is XRPD patternof adipate Form A after test);

FIG. 30 DVS curve of maleate Form A (0-95% RH cycle);

FIG. 31 XRPD overlay of maleate Form A before and after hygroscopicitytest (a is XRPD pattern of maleate Form A before test, b is XRPD patternof maleate Form A after test);

FIG. 32 DVS curve of glycollate Form A (0-95% RH cycle);

FIG. 33 XRPD overlay of glycollate Form A before and afterhygroscopicity test (a is XRPD pattern of glycollate Form A before test,b is XRPD pattern of glycollate Form A after test);

FIG. 34 DVS curve of mono-succinate non-hydrate form in CN103201275A(0-95% RH cycle);

FIG. 35 XRPD overlay of mono-succinate non-hydrate form in patentCN103201275A before and after hygroscopicity test (the pattern below isXRPD pattern before test, the pattern above is XRPD pattern after test,form changed);

DETAILED DESCRIPTION OF THE INVENTION

The present invention will be further explained by the specificembodiments, but are not intended to limit the scope of the presentinvention. The skilled in the art can make improvements to the processand the used instruments within the scope of the claims, and thoseimprovements should be considered as falling into the scope of thepresent invention. Accordingly, the protective scope of the presentinvention patent should be defined by the appended claims.

The abbreviations used in the invention are explained as follows:

XRPD: X-ray Powder Diffraction

DSC: Differential Scanning calorimetry

TGA: Thermal Gravimetric Analysis

DVS: Dynamic Vapor Sorption

¹H NMR: ¹H Nuclear Magnetic Resonance

X-ray powder diffraction pattern in the present invention was acquiredby a Panalytical Empyrean X-ray powder diffractometer. The parameters ofthe X-ray powder diffraction method of the present invention were asfollows:

X-ray Reflection: Cu, Kα

Kα1 (Å): 1.540598; Kα2 (Å): 1.544426

Kα2/Kα1 intensity ratio: 0.50

Voltage: 45 (kV)

Current: 40 (mA)

Scan range: from 3.0 degree to 40.0 degree

The pattern of differential scanning calorimetry (DSC) in the presentinvention was acquired by a TA Q2000. The parameters of the differentialscanning calorimetry (DSC) method of the present invention were asfollow:

Heating rate: 10° C./min

Purge gas: nitrogen.

The pattern of thermal gravimetric analysis (TGA) in the presentinvention was acquired by a TA Q5000. The parameters of the thermalgravimetric analysis (TGA) method of the present invention were asfollow:

Heating rate: 10° C./min;

Purge gas: nitrogen.

Dynamic Vapor Sorption (DVS) was measured via a SMS (Surface MeasurementSystems) DVS Intrinsic. Typical Parameters for DVS test are listedbelow.

Temperature: 25° C.

Gas and flow rate: N2, 200 mL/min

dm/dt: 0.002%/min

RH range: 0% RH to 95% RH

EXAMPLE 1

Process for Preparing Hemi-Succinate Form A:

10.5 mg of compound I freebase was added into ethanol, and 3.0 mg ofsuccinic acid was added, then stirred at room temperature for 12 hoursuntil solids precipitate out. Hemi-succinate Form A was analyzed byXRPD, DSC, TGA and ¹H NMR. The XRPD data of the hemi-succinate Form Aproduced in this example is listed in Table 1. The DSC data shows anendothermic peak at 180° C. (onset temperature). The TGA data shows12.5% weight loss up to 118° C. The XRPD pattern is displayed in FIG. 1,the ¹H NMR spectrum is displayed in FIG. 2.

¹H NMR data of hemi-succinate Form A produced in this example is shownas following: ¹H NMR (400 MHz, DMSO) δ 9.29 (s, 1H), 8.76 (s, 1H), 8.16(d, J=9.0 Hz, 1H), 8.00 (d, J=2.9 Hz, 1H), 7.44 (dd, J=9.2, 3.0 Hz, 1H),6.60 (s, 1H), 3.15-2.93 (m, 14H), 2.32 (s, 2H), 1.98 (s, 4H), 1.65 (s,2H).

TABLE 1 2theta d spacing Intensity % 4.69 18.84 71.02 8.87 9.97 23.859.45 9.36 16.73 10.64 8.32 41.57 12.96 6.83 23.59 14.24 6.22 63.46 15.735.63 7.33 16.17 5.48 18.29 16.47 5.38 6.10 17.83 4.97 34.08 18.26 4.8620.26 18.47 4.80 37.64 19.04 4.66 51.36 19.89 4.46 34.73 20.04 4.4340.63 20.70 4.29 11.44 21.29 4.17 100.00 21.57 4.12 17.28 22.04 4.0340.12 22.20 4.00 40.13 22.61 3.93 7.68 23.88 3.73 78.70 24.32 3.66 25.9225.33 3.52 7.29 26.20 3.40 6.47 26.72 3.34 24.03 27.83 3.21 7.19 28.783.10 11.72 31.42 2.85 5.46 32.50 2.76 2.50 33.72 2.66 5.12 38.65 2.332.58

EXAMPLE 2

Process for Preparing of Hemi-Succinate Form A:

10.2 mg of compound I freebase was added into tetrahydrofuran, and 2.8mg of succinic acid was added, then stirred at room temperature for 12hours until solids precipitate out. The XRPD data of the hemi-succinateForm A produced in this example is listed in Table 2.

TABLE 2 2theta d spacing Intensity % 4.70 18.81 100.00 6.14 14.40 13.178.87 9.96 12.08 10.65 8.31 24.56 12.32 7.18 7.43 12.97 6.82 23.94 14.286.20 67.09 16.18 5.48 21.53 17.87 4.96 34.36 18.49 4.80 56.14 19.10 4.6560.98 20.08 4.42 49.94 20.72 4.29 19.08 21.32 4.17 60.86 21.56 4.1228.62 22.04 4.03 54.18 22.21 4.00 43.36 22.64 3.93 11.04 23.49 3.7916.24 23.89 3.72 87.76 24.36 3.65 31.70 25.31 3.52 16.68 26.24 3.4013.04 26.75 3.33 33.25 27.87 3.20 16.12 28.82 3.10 14.01 30.60 2.92 5.0031.43 2.85 12.77 32.60 2.75 5.05 33.72 2.66 7.39 35.71 2.51 5.65 38.622.33 4.11

EXAMPLE 3

Stability of Hemi-Succinate Form A at High Humidity:

About 10.0 mg of hemi-succinate Form A was analyzed by DVS. The solidform was tested by XRPD before and after DVS. The result showed that ithas a 3.5% weight gain at 80% RH, which is slightly hygroscopic. The DVScurve was displayed in FIG. 3, the XRPD overlay pattern is displayed inFIG. 4.

About hygroscopicity characterization description and definition ofhygroscopicity (Chinese Pharmacopoeia 2010 edition appendix XIXJ Drughygroscopic test guidelines, test at 25° C.+/−1° C., 80% RelativeHumidity)

-   -   deliquescent: sufficient water is absorbed to form a liquid;    -   very hygroscopic: increase in mass is equal to or greater than        15 percent;    -   hygroscopic: increase in mass is less than 15 percent and equal        to or greater than 2 percent;    -   slightly hygroscopic: increase in mass is less than 2 percent        and equal to or greater than 0.2 percent.    -   no or almost no hygroscopic: increase in mass is less than 0.2%

EXAMPLE 4

Conversion Relationship Between Hemi-Succinate Form A and Mono-SuccinateNon-Hydrate Form in Patent CN103201275A:

About 10 mg of the non-hydrate form in patent CN103201275A as startingform was added in different solvents or mixed solvents (v/v), thenstirred at 5-50° C. for about 48 hours. Finally, the starting formconverted to hemi-succinate Form A. Solvents used in this example isshown in table 3.

TABLE 3 Solvent/Mixed solvent NO. Starting Form (v/v) Final Form 1mono-succinate in Ethyl acetate hemi-succinate patent CN103201275A FormA 2 mono-succinate in Ethanol:water = 20:1 hemi-succinate patentCN103201275A Form A 3 mono-succinate in Acetone:water = 20:1hemi-succinate patent CN103201275A Form A 4 mono-succinate inTetrahydrofuran:water = hemi-succinate patent CN103201275A 20:1 Form A

EXAMPLE 5

Process for Preparing Mono-Succinate Form I:

30.7 mg of the non-hydrate form (prepared according to patentCN103201275A) was added into 2.2 mL of acetonitrile/methanol (v/v=10/1),then stirred at 50° C. for 48 hours, until solids precipitate out.

The XRPD data of the mono-succinate Form I produced in this example islisted in Table 4 and the XRPD pattern was displayed in FIG. 5.

TABLE 4 2theta d spacing Intensity % 6.83 12.95 7.85 7.82 11.30 16.0611.24 7.88 3.49 11.88 7.45 53.46 12.46 7.11 5.71 13.06 6.78 19.20 13.316.65 31.18 14.01 6.32 16.85 15.71 5.64 20.66 16.29 5.44 6.07 16.70 5.3121.23 17.81 4.98 71.41 18.63 4.76 18.63 19.35 4.59 31.47 19.69 4.51 8.4720.10 4.42 32.20 20.47 4.34 43.73 20.64 4.30 100.00 21.23 4.19 16.2621.72 4.09 6.12 22.74 3.91 61.23 23.12 3.85 15.91 23.34 3.81 6.29 23.893.73 5.74 24.43 3.64 66.75 25.11 3.55 4.94 25.80 3.45 7.99 26.27 3.3923.61 27.61 3.23 8.03 27.99 3.19 4.23 28.44 3.14 82.46 28.52 3.14 46.4429.18 3.06 14.13 29.74 3.00 3.00 30.04 2.97 3.93 30.82 2.90 3.98 31.162.87 3.74 31.59 2.83 4.01 32.27 2.77 3.61 33.08 2.71 7.40 34.21 2.622.30 36.23 2.48 3.61 37.35 2.41 1.65 38.69 2.33 1.28

The ¹H NMR spectrum of the mono-succinate Form I produced in thisexample is displayed in FIG. 6. ¹H NMR data is shown as following:

¹H NMR (400 MHz, DMSO) δ 9.33 (s, 1H), 8.76 (s, 1H), 8.16 (d, J=9.1 Hz,1H), 8.00 (d, J=2.9 Hz, 1H), 7.45 (dd, J=9.1, 3.0 Hz, 1H), 6.60 (s, 1H),4.79-4.68 (m, 1H), 3.16-3.00 (m, 14H), 2.34 (s, 4H), 1.98 (s, 4H), 1.64(d, J=5.5 Hz, 2H). ¹H NMR results show that Form I is a mono-succinateof compound I.

DSC curve of mono-succinate Form I was displayed in FIG. 7. Form I is ananhydrate, the DSC data showed an endothermic peak at 197° C. (onsettemperature).

TGA curve of mono-succinate Form I was displayed in FIG. 8. The TGA datashowed 2.0% weight loss up to 178° C.

EXAMPLE 6

Process for Preparing Mono-Succinate Form I:

3.1 mg of the non-hydrate form (prepared according to patentCN103201275A) was added into 0.5 mL of ethanol/n-heptane (v/v=4/1), thenstirred at room temperature for 48 hours until solids precipitate out.The XRPD data of the mono-succinate Form I produced in this example islisted in Table 5.

TABLE 5 2theta d spacing Intensity % 6.83 12.95 11.44 7.84 11.28 26.2411.90 7.44 88.02 13.09 6.77 21.52 13.33 6.64 35.24 14.02 6.31 15.8515.74 5.63 23.50 16.71 5.31 18.54 17.83 4.98 76.15 18.64 4.76 22.5119.37 4.58 31.64 20.11 4.41 28.17 20.65 4.30 100.00 21.26 4.18 14.8222.22 4.00 5.89 22.76 3.91 63.69 23.15 3.84 17.57 24.44 3.64 71.31 25.123.54 7.95 25.81 3.45 8.77 26.29 3.39 19.56 27.59 3.23 5.79 28.16 3.171.30 29.20 3.06 17.50 30.11 2.97 4.35 30.85 2.90 5.16 32.30 2.77 4.9533.11 2.71 7.22 34.25 2.62 2.79 36.26 2.48 3.54

EXAMPLE 7

Stability of Mono-Succinate Form I of the Present Invention andNon-Hydrate Form in Patent CN103201275A at High Humidity:

About 10 mg of mono-succinate Form I of the present invention andnon-hydrate form in patent CN103201275A were analyzed by DVS. The solidwas tested by XRPD before and after hygroscopicity test. The DVS curvewas displayed in FIG. 9, the XRPD overlay pattern before and afterhygroscopicity test is displayed in FIG. 10. The result showed that itadsorbed 1.7% water at 25° C./90% RH, and Form I of the presentinvention did not change after hygroscopicity test. It indicated thatForm I of the present invention was stable at high humidity. Accordingto the data in patent CN103201275A, the non-hydrate form adsorbed 2.0%water at 25° C./90% RH, and 7.35% of the non-hydrate form converted tohydrate form. Furthermore, 0.52% of the non-hydrate form in patentCN103201275A converted to hydrate form at 25° C./80% RH.

As shown in FIG. 9 and FIG. 10, Form I of the present invention adsorbed4.0% water at 25° C./95% RH, and Form I of the present invention did notchange after test. As shown in FIG. 11, the non-hydrate form adsorbed18.3% water at 25° C./95% RH, and the solid form changed after test. Theresults indicated that Form I of the present invention was stable athigh humidity and the solid form did not change, while the non-hydrateform in patent CN103201275A was not stable as it would easily havecrystal transformation at high humidity.

EXAMPLE 8

Stability of Form I of the Present Invention and Mono-SuccinateNon-Hydrate Form in Patent CN103201275A in Different Temperatures:

About 10 mg of the non-hydrate form in patent CN103201275A as startingform was added in different solvents or mixed solvents (v/v), thenstirred at 5-50° C. for about 48 hours. Finally, the starting formconverted to Form I. Solvents and temperature used in this example is inthe following in table 6.

TABLE 6 Solvent/Mixed NO. Starting Form Temperature solvent (v/v) FinalForm 1 mono-succinate in patent RT Ethanol:n- mono-succinateCN103201275A heptane = 4:1 Form I 2 mono-succinate in patent  5° C.Ethanol:n- mono-succinate CN103201275A heptane = 4:1 Form I 3mono-succinate in patent 50° C. Acetonitrile:water = mono-succinateCN103201275A 20:1 Form I 4 mono-succinate in patent 50° C. THF:Methanol= mono-succinate CN103201275A 20:1 Form I

EXAMPLE 9

Process for Preparing Adipate:

200 mg of compound I freebase powder was added into 10.0 mL ofacetone/water (v/v=19/1), and 68 mg of adipic acid was added to thesolution, then stirred at room temperature, the solid was obtained. The¹H NMR spectrum is displayed in FIG. 22.

¹H NMR data of adipate Form A produced in this example is shown asfollowing:

¹H NMR (400 MHz, DMSO) δ 9.31 (s, 1H), 8.76 (s, 1H), 8.15 (d, J=9.1 Hz,1H), 7.99 (d, J=2.8 Hz, 1H), 7.42 (dd, J=9.1, 3.0 Hz, 1H), 6.60 (s, 1H),4.78-4.67 (m, 1H), 3.06 (d, J=4.9 Hz, 10H), 2.95-2.82 (m, 4H), 2.48-2.38(m, 2H), 2.25-2.09 (m, 4H), 1.98 (s, 4H), 1.64 (d, J=4.9 Hz, 2H),1.54-1.38 (m, 4H).

The result shows the solid is adipate Form A. The XRPD data of theadipate Form A produced in this example are listed in Table 7. The XRPDpattern is displayed in FIG. 13, the DSC curve is displayed in FIG. 14,the TGA curve is displayed in FIG. 15.

TABLE 7 2theta d spacing Intensity % 4.79 18.45 17.56 9.64 9.17 11.1710.71 8.26 3.80 11.41 7.75 3.13 12.16 7.28 8.75 12.38 7.15 11.40 12.567.05 15.09 12.89 6.87 9.06 13.97 6.34 24.14 14.43 6.14 14.43 15.07 5.883.07 15.64 5.66 6.31 16.09 5.51 20.51 16.96 5.23 13.46 17.30 5.12 8.9617.80 4.98 17.12 18.03 4.92 17.83 18.18 4.88 13.71 18.49 4.80 7.77 19.154.64 83.86 19.39 4.58 53.64 19.80 4.48 17.69 20.34 4.37 8.25 20.58 4.324.02 21.24 4.18 8.19 21.43 4.15 7.31 21.81 4.08 7.02 22.20 4.00 100.0023.32 3.81 4.96 24.34 3.66 6.11 24.62 3.62 8.83 24.91 3.58 29.19 25.343.51 14.68 25.72 3.46 5.59 25.90 3.44 6.02 26.41 3.37 3.21 27.09 3.297.60 27.60 3.23 5.29 28.24 3.16 12.15 29.11 3.07 3.52 29.40 3.04 10.5229.48 3.03 10.46 29.84 2.99 1.80 30.26 2.95 1.39 31.39 2.85 4.89 32.092.79 8.61 32.39 2.76 6.51 32.56 2.75 5.71 33.53 2.67 2.54 34.28 2.623.59

EXAMPLE 10

Process for Preparing Adipate:

10.3 mg of compound I freebase powder was added into 0.4 mL ofacetone/water (v/v=19/1), and 3.9 mg of adipic acid was added to thesolution, then stirred at room temperature, the solid was obtained.

The solid is adipate Form A after analysis. XRPD data is displayed intable 8

TABLE 8 2theta d spacing Intensity % 4.85 18.22 100.00 9.67 9.15 19.1212.37 7.16 11.38 12.58 7.04 12.33 13.97 6.34 25.90 14.47 6.12 13.9016.09 5.51 30.88 16.94 5.23 7.33 17.82 4.98 13.28 18.11 4.90 8.57 19.114.64 52.72 19.36 4.59 63.62 19.79 4.49 9.60 20.34 4.37 8.80 21.38 4.165.23 22.20 4.00 83.50 24.94 3.57 18.93 25.34 3.51 9.62 28.24 3.16 5.7629.46 3.03 6.03 31.41 2.85 3.22 32.13 2.79 5.49 32.50 2.76 3.51 35.542.53 2.90

EXAMPLE 11

Process for Preparing Maleate:

200.63 mg of compound I freebase powder was added into 10.0 mL ofacetone/water (v/v=19/1), and 56 mg of maleic acid was added to thesolution, then stirred at room temperature, the solid was obtained, The¹H NMR spectrum is displayed in FIG. 23.

¹H NMR data of the maleate Form A produced in this example are shown asfollowing:

¹H NMR (400 MHz, DMSO) δ 9.49 (s, 1H), 8.77 (s, 1H), 8.20 (d, J=9.1 Hz,1H), 8.07 (d, J=2.8 Hz, 1H), 7.52 (dd, J=9.1, 2.8 Hz, 1H), 6.62 (s, 1H),6.04 (s, 2H), 4.80-4.66 (m, 1H), 3.34 (d, J=5.6 Hz, 4H), 3.28 (d, J=5.3Hz, 4H), 3.06 (s, 6H), 2.48-2.35 (m, 2H), 1.98 (s, 4H), 1.65 (d, J=5.3Hz, 2H).

The result shows the solid is maleate Form A. The XRPD data of themaleate Form A is listed in Table 9. The XRPD pattern is displayed inFIG. 16, the DSC curve is displayed in FIG. 17, the TGA curve isdisplayed in FIG. 18.

TABLE 9 2theta d spacing Intensity % 8.52 10.38 30.01 10.64 8.31 16.4310.99 8.05 3.98 11.35 7.80 13.13 12.35 7.17 21.68 12.59 7.03 15.42 14.256.22 11.75 14.90 5.95 56.02 15.93 5.56 28.44 16.53 5.36 36.61 17.10 5.1929.08 17.67 5.02 8.92 18.05 4.91 23.39 18.55 4.78 100.00 18.94 4.6913.24 19.22 4.62 11.29 19.90 4.46 94.35 20.35 4.36 19.29 21.13 4.2025.99 21.78 4.08 36.87 21.93 4.05 35.06 22.31 3.98 13.67 22.85 3.8914.35 23.83 3.73 15.82 24.47 3.64 38.79 25.35 3.51 12.11 25.87 3.4423.90 27.07 3.29 21.86 27.42 3.25 27.86 27.79 3.21 14.66 28.88 3.09 9.8829.26 3.05 30.60 30.14 2.97 9.28 30.80 2.90 8.50 31.49 2.84 13.38 32.822.73 4.33 36.00 2.49 1.81 37.70 2.39 3.06 38.44 2.34 1.34

EXAMPLE 12

Process for Preparing Maleate:

10.3 mg of compound I freebase powder was added into 0.4 mL of acetone,and 2.8 mg of maleic acid was added to the solution, then stirred atroom temperature, the solid was obtained.

The solid is maleate Form A after analysis. XRPD data is displayed intable 10.

TABLE 10 2theta d spacing Intensity % 5.41 16.33 37.33 7.81 11.32 33.488.54 10.36 37.61 10.65 8.31 22.54 12.35 7.17 29.59 14.90 5.95 71.3315.95 5.56 33.17 16.54 5.36 35.61 17.10 5.18 33.04 18.07 4.91 26.1218.53 4.79 100.00 19.92 4.46 97.61 21.13 4.21 22.94 21.88 4.06 42.2924.51 3.63 32.45 25.95 3.43 15.94 27.29 3.27 21.96 29.28 3.05 26.9831.58 2.83 7.08

EXAMPLE 13

Process for Preparing Glycollate:

199.0 mg of compound I freebase powder was added into 10.0 mL ofacetone/water (v/v=19/1), and 34.0 mg of glycolic acid was added to thesolution, then stirred at room temperature, the solid was obtained. TheH¹NMR spectrum is displayed in FIG. 24.

¹H NMR data of glycollate Form A produced in this example is shown asfollowing:

¹H NMR (400 MHz, DMSO) δ 9.09 (d, J=10.7 Hz, 1H), 8.53 (s, 1H), 7.93 (d,J=9.1 Hz, 1H), 7.78 (d, J=2.9 Hz, 1H), 7.21 (dd, J=9.1, 2.9 Hz, 1H),6.37 (s, 1H), 4.55-4.45 (m, 1H), 3.54 (s, 2H), 2.95-2.87 (m, 4H), 2.83(s, 6H), 2.79-2.74 (m, 4H), 2.24-2.17 (m, 2H), 1.75 (s, 4H), 1.41 (d,J=5.0 Hz, 2H).

The result shows the solid is glycollate Form A. The XRPD data of theglycollate Form A is listed in Table 11. The XRPD pattern is displayedin FIG. 19, the DSC curve is displayed in FIG. 20, the TGA curve isdisplayed in FIG. 21,

TABLE 11 2theta d spacing Intensity % 5.13 17.22 9.69 7.61 11.61 5.7210.06 8.79 26.70 10.29 8.60 9.05 11.08 7.99 8.79 12.43 7.12 27.49 13.266.68 23.74 14.90 5.95 10.46 15.42 5.75 8.40 16.82 5.27 21.30 17.84 4.979.39 18.30 4.85 10.54 19.46 4.56 74.36 21.08 4.21 21.65 21.28 4.17100.00 21.79 4.08 30.27 22.55 3.94 4.22 22.95 3.87 7.78 23.26 3.82 31.8223.90 3.72 11.86 25.31 3.52 7.49 25.92 3.44 5.24 26.72 3.34 7.72 27.043.30 1.06 27.30 3.27 6.40 28.49 3.13 7.19 28.92 3.09 2.10 30.10 2.974.36 32.21 2.78 2.03 33.47 2.68 4.76 34.04 2.63 3.14 34.57 2.59 2.9235.49 2.53 1.49 36.61 2.45 1.93 37.98 2.37 2.93

EXAMPLE 14

Process for Preparing Glycollate:

10.3 mg of compound I freebase powder was added into 0.4 mL ofacetone/water (v/v=19/1), and 4.2 mg of glycolic acid was added to thesolution, then stirred at room temperature, the solid was obtained.

The solid is glycollate Form A after analysis. XRPD data is displayed intable 12.

TABLE 12 2theta d spacing Intensity % 5.11 17.30 21.55 10.03 8.82 35.4011.03 8.02 5.25 12.40 7.14 33.03 13.23 6.69 26.09 14.90 5.95 8.26 15.405.75 4.84 16.77 5.29 17.99 17.83 4.98 7.39 18.28 4.85 6.80 19.45 4.5687.19 20.65 4.30 4.90 21.24 4.18 100.00 21.76 4.08 37.62 22.94 3.88 9.0323.22 3.83 29.44 23.91 3.72 14.57 25.29 3.52 6.02 25.90 3.44 5.66 26.733.34 8.95 27.27 3.27 6.00 28.49 3.13 8.21 28.94 3.09 2.58 30.09 2.975.59 31.16 2.87 3.39 33.48 2.68 5.30 34.10 2.63 3.59 34.56 2.60 3.8835.34 2.54 2.36 36.59 2.46 2.50 37.95 2.37 3.36

EXAMPLE 15

Stability of Crystalline Salt Forms of Compound I:

Two samples prepared by example 9, example 11 and example 13 were storedfor 30 days under 25° C./60% RH and 40° C./75% RH. The samples beforeand after storage were tested by XRPD, the results were summarized intable 13.

TABLE 13 Storage Storage HPLC Starting Form Condition Time Solid FormPurity (%) Adipate Form A 25° C./60% RH 30 days No Change 99.1 (FIG.25a) (FIG. 25b) Adipate Form A 40° C./75% RH 30 days No Change 98.9(FIG. 25a) (FIG. 25c) Maleate Form A 25° C./60% RH 30 days No Change99.3 (FIG. 26a) (FIG. 26b) Maleate Form A 40° C./75% RH 30 days NoChange 99.3 (FIG. 26a) (FIG. 26c) Glycollate TypeA 25° C./60% RH 30 daysNo Change 99.7 (FIG. 27a) (FIG. 27b) Glycollate TypeA 40° C./75% RH 30days No Change 99.6 (FIG. 27a) (FIG. 27c)

The results indicate that the crystalline salt forms of compound I didnot change after 30 days. In conclusion, the result proves excellentstability of crystalline salt forms of compound I.

EXAMPLE 16

Stability of Salts in Present Invention and Mono-Succinate in PatentCN103201275A at High Humidity:

About 10 mg of adipate Form A, maleate Form A, glycollate Form A andmono-succinate non-hydrate form in patent CN103201275A were analyzed byDVS. The solid was tested by XRPD before and after hygroscopicity test.The results were displayed in table 14, the DVS curve of adipate Form Awas displayed in FIG. 28, the XRPD overlay pattern is displayed in FIG.29. The DVS curve of maleate Form A was displayed in FIG. 30, the XRPDoverlay pattern was displayed in FIG. 31. The DVS curve of glycollateForm A was displayed in FIG. 32, the XRPD overlay pattern was displayedin FIG. 33. The DVS curve of non-hydrate form in patent CN103201275A wasdisplayed in FIG. 34, the XRPD overlay pattern was displayed in FIG. 35(the pattern below is before DVS, the pattern above is after DVS).

TABLE 14 Water absorption at Starting Form 25° C./95% RH Change afterDVS adipate Form A 0.7 No change maleate Form A 3.4 No change glycollateForm A 1.2 No change mono-succinate 18.3 Change non-hydrate form inpatent CN103201275A

The results indicated that adipate Form A, maleate Form A and glycollateForm A were stable at high humidity and the solid form did not change.While the non-hydrate form in patent CN103201275A was not stable as itchanged in high humidity.

The examples described above are only for illustrating the technicalconcepts and features of the present invention, and intended to makethose skilled in the art being able to understand the present inventionand thereby implement it, and should not be concluded to limit theprotective scope of this invention. Any equivalent variations ormodifications according to the spirit of the present invention should becovered by the protective scope of the present invention.

The invention claimed is:
 1. A crystalline mono-succinate Form I ofcompound I,

wherein the X-ray powder diffraction pattern of the crystalline Form Ishows characteristic peaks at 2theta values of 11.9°±0.2°, 19.4°±0.2°,20.6°±0.2°.
 2. The crystalline Form I according to claim 1, wherein theX-ray powder diffraction pattern of the crystalline Form I shows one ortwo or three of the characteristic peaks at 2theta values of 22.7°±0.2°,24.4°±0.2°, 26.3°±0.2°.
 3. The crystalline Form I according to claim 2,wherein the X-ray powder diffraction pattern of the crystalline Form Ishows characteristic peaks at 2theta values of 22.7°±0.2°, 24.4°±0.2°,26.3°±0.2°.
 4. The crystalline Form I according to claim 1, wherein theX-ray powder diffraction pattern of the crystalline Form I shows one ortwo or three of the characteristic peaks at 2theta values of 7.8°±0.2°,15.7°±0.2°, 16.7°±0.2°.
 5. The crystalline Form I according to claim 4,wherein the X-ray powder diffraction pattern of the crystalline Form Ishows characteristic peaks at 2theta values of 7.8°±0.2°, 15.7°±0.2°,16.7°±0.2°.
 6. The crystalline Form I according to claim 1, wherein saidForm I has an X-ray powder diffraction pattern substantially as depictedin FIG.
 5. 7. A process for preparing the crystalline Form I accordingto claim 1, comprising: dissolving compound I mono-succinate in a mixedsolvent of alcohols and one or more solvents selected from the groupconsisting of alkyl nitriles and alkanes, or dissolving compound Imono-succinate in a mixture of water and one or more solvents selectedfrom the group consisting of alkyl nitriles, to form a mixture; andstirring the mixture until solids precipitate out.
 8. The processaccording to claim 7, wherein said alcohol in mixed solvent is methanol,ethanol or a combination thereof, said alkyl nitrile in mixed solvent isacetonitrile, said alkane in mixed solvent is n-heptane.
 9. The processaccording to claim 8, wherein said mixed solvents are acetonitrile andmethanol, ethanol and n-heptane or acetonitrile and water.
 10. Apharmaceutical composition comprising a therapeutically effective amountof the mono-succinate Form I according to claim 1 and pharmaceuticalacceptable carrier.